Primer system and applications thereof

A technology of nucleotide sequences and expression vectors, applied in the primer system and its application fields, can solve the problems of unfavorable environment, high cost, waste of living materials, etc.

Inactive Publication Date: 2015-05-20
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Since the amount of AMPs naturally present in most organisms is very small, the amount of AMPs obtained by this method is also very small, and it will lead to a waste of a large amount of living materials, which is not good for the enviro

Method used

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  • Primer system and applications thereof
  • Primer system and applications thereof
  • Primer system and applications thereof

Examples

Experimental program
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Embodiment 1

[0033] This example is used to illustrate the acquisition of CFB

[0034] 1. Overlap PCR to obtain the target gene

[0035] Synthesize the following four primers:

[0036] P1: Add a restriction endonuclease KpnI enzyme cutting site (GGTACC) and a protection base CGG to the 5' end of primer 1 shown in SEQ ID No: 1;

[0037] P2; as shown in SEQ ID No: 2;

[0038] P3; as shown in SEQ ID No: 3;

[0039] P4; at the 5' end of primer 4 shown in SEQ ID No: 4, a restriction endonuclease XhoI restriction site (CTCGAG) and a protective base CGG are added;

[0040] PCR reaction system: (Pfμmixtμre was purchased from Bao Biological Engineering (Dalian) Co., Ltd.)

[0041]

[0042]

[0043] PCR reaction program: pre-denaturation at 95°C for 5 minutes, followed by denaturation at 95°C for 45s, annealing at 60°C for 30s, extension at 72°C for 30s, and 35 cycles; after the cycle, extend at 72°C for 10 minutes and cool to 4°C.

[0044]Use the Zymoclean Gel DNA Recovery Kit (ZYMO RESE...

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Abstract

The invention discloses a primer system. The primer system comprises a primer 1, a primer 2, a primer 3 and a primer 4, wherein the nucleotide sequence of the primer 1 is shown in SEQ ID No:1, the nucleotide sequence of the primer 2 is shown in SEQ ID No:2, the nucleotide sequence of the primer 3 is shown in SEQ ID No:3, and the nucleotide sequence of the primer 4 is shown in SEQ ID No:4. The invention also discloses an expression vector and a transgenic cell and applications of the expression and transgenic cell. Through the technical scheme above, the heterologous in vitro expression of krait antibacterial peptides CBF is realized for the first time, and a purpose of rapidly obtaining large quantities of CBF in a low-cost mode is successfully achieved, thereby providing a theoretical basis for the large-scale development and utilization of CBF.

Description

technical field [0001] The present invention relates to a primer system and its application, in particular to a primer system for amplifying the antimicrobial peptide Cathelicidin-BF (CFB), which contains an expression vector for the amplified product of the primer system, containing the The transgenic cell expressing the vector, and the application of the primer system, the expression vector and the transgenic cell in antibacterial in vitro and / or in the preparation of bacteriostatic medicine. Background technique [0002] In recent years, the problem of microbial resistance due to the abuse of antibiotics has become more and more serious, and new drug-resistant strains and drug-resistant incidents have been reported continuously. In the past 40 years, only three new antibiotics have been marketed, and all of them are used to treat Gram-positive bacterial infections, so the development of new antibiotic alternatives is imminent. Antimicrobial peptides (AMPs), as a kind of ...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/70C12N1/21A61K48/00A61K35/74A61P31/00C12R1/19
Inventor 任发政赵亮姜鹭刘松玲郭慧媛王一博
Owner CHINA AGRI UNIV
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